A long-term field experiment on soil fertility and fertilizer efficiency in flue-cured tobacco cultivation was established in 2004 at Zunyi， Guizhou. Relying on this long-term field experiment， the changes in tobacco leaf yield and quality， fertilizer agronomic efficiency， and nitrogen (N) and enzyme activity in the soil under the different tobacco planting (continuous tobacco cultivation， tobacco-maize rotation) and different fertilization (chemical fertilizer alone， combined application of manure plus chemical fertilizers) were investigated in 2017， 2019， and 2021. The objectives were to realize the relationships between tobacco agronomic performances and cultivation models or fertilization practices and to provide a scientific basis for N application in flue-curried tobacco cultivation. Compared with continuous tobacco cultivation， tobaccomaize rotation was increased the leaf yield by 31.4%， the proportion of middle and top grade leaves by 6.2%， leaf economic output by 37.2%， fertilizer agronomic efficiency by 35.9%， and fertilizer economic utilization efficiency by 31.4%. These results indicated that tobacco-maize rotation was beneficial to the improvement of the tobacco leaf yield， quality. In the continuous cropping mode， combined application of manure plus chemical fertilizer application (MCF) increased the abovementioned parameters by 24.7%， 9.1%， 28.9%， and 24.8%， respectively， compared with chemical fertilizer alone. In the tobacco-maize rotation soil， MCF also increased the leaf yield and economic output in two of the three seasons and generally improved the leaf quality. These phenomena are closely related to autotoxicity alleviation， increase in total and alkaline hydrolyzed N， ammonium N, nitrate N, enzyme stimulation (protease， urease， nitrate and nitrite reductase)， N transformation promotion， nitrogen supply improvement， and nitrate N enhancement by MCF in the tobacco-maize rotation soil. Taken together， it is worth advocating tobacco-corn rotation and chemical fertilizer combined with organic fertilizer in the northern Guizhou tobacco region.